Zhang Zhe, Chen Cheng-Zhen, Xu Ming-Qiang, Zhang Li-Qun, Liu Jian-Bo, Gao Yan, Jiang Hao, Yuan Bao, Zhang Jia-Bao
Department of Laboratory Animals, College of Animal Sciences, Jilin University, Changchun, 130062, Jilin, PR China.
Department of Obstetrics and Gynecology, The First Hospital of Jilin University, Changchun, 130012, Jilin, PR China.
Theriogenology. 2019 Jan 1;123:45-53. doi: 10.1016/j.theriogenology.2018.09.020. Epub 2018 Sep 26.
The regulatory role of microRNAs (miRNAs) has been explored in ovarian cells, and the effects of miRNAs on gonadal development, apoptosis, ovulation, and steroid production have been reported. In this study, we analyzed the effects of follicle stimulating hormone (FSH) on miR-31 and miR-143 expression levels in bovine granulosa cells (GCs). Our results demonstrated that the FSH receptor (FSHR) is a common target gene of miR-31 and miR-143 in bovine GCs. We further analyzed the roles of miR-31 and miR-143 in bovine GCs by transfecting miR-31 and miR-143 mimics and inhibitors. The Western blot and RT-PCR results showed that miR-31 and miR-143 reduced the mRNA and protein expression levels of FSHR. Moreover, miR-31 overexpression decreased the secretion of progesterone (P4), and miR-143 overexpression decreased both the synthesis of P4 and the secretion of estrogen (E2). In contrast, miR-31 inhibition increased the secretion of progesterone (P4), and miR-143 inhibition increased both the synthesis of P4 and the secretion of E2. Finally, we analyzed the possible effects of miR-31 and miR-143 on bovine GC apoptosis. The results showed that transfection with miR-31 and miR-143 mimics promoted GC apoptosis and that miR-143 and miR-31 inhibition reduced the rate of apoptosis in bovine GCs. Taken together, our results indicate that miR-31 and miR-143 decrease steroid hormone synthesis and inhibit bovine GC apoptosis by targeting FSHR.
微小RNA(miRNA)在卵巢细胞中的调控作用已得到研究,并且已有报道称miRNA对性腺发育、细胞凋亡、排卵和类固醇生成有影响。在本研究中,我们分析了促卵泡激素(FSH)对牛颗粒细胞(GCs)中miR-31和miR-143表达水平的影响。我们的结果表明,FSH受体(FSHR)是牛GCs中miR-31和miR-143的共同靶基因。我们通过转染miR-31和miR-143模拟物及抑制剂,进一步分析了miR-31和miR-143在牛GCs中的作用。蛋白质免疫印迹和逆转录-聚合酶链反应结果显示,miR-31和miR-143降低了FSHR的mRNA和蛋白质表达水平。此外,miR-31过表达降低了孕酮(P4)的分泌,miR-143过表达降低了P4的合成及雌激素(E2)的分泌。相反,抑制miR-31增加了P4的分泌,抑制miR-143增加了P4的合成及E2的分泌。最后,我们分析了miR-31和miR-143对牛GC凋亡的可能影响。结果显示,转染miR-31和miR-143模拟物可促进GC凋亡,而抑制miR-143和miR-31可降低牛GCs的凋亡率。综上所述,我们的结果表明,miR-31和miR-143通过靶向FSHR降低类固醇激素合成并抑制牛GC凋亡。